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Post-operative epidural fibrosis is a biological response after laminectomy that may lead to clinical symptoms, such as radicular pain. An ideal material for prevention of epidural fibrosis should be able to inhibit fibroblast adhesions and reduce formation of scar tissue. An injectable hydrogel would be the material of choice for this purpose, since it could fill an irregular surgical defect completely, gelate in situ and be delivered in a minimally-invasive manner. The objective of this study was to evaluate, in vitro and in vivo, the cytocompatibility and anti-adhesive effect of an oxidised hyaluronic acid/adipic acid dihydrazide (oxi-HA/ADH) hydrogel. Different cell types present in the spine were used to test the cytocompatibility of the hydrogel. The hydrogel extraction medium had no deleterious effects on neural cells (PC-12), but reduced fibroblasts viability (NIH/3T3). Although the hydrogel did not change the release of lactate dehydrogenase from myoblasts (C2C12) and Schwann cells (RSC96), the extraction medium concentration slightly affected the mitochondrial activity of these two cell types. qPCR showed that the hydrogel down-regulated S100a and P4hb expression in NIH/3T3 cells, supporting the hypothesis that the hydrogel might inhibit fibroblast activity. The animal study showed a reduction of scar tissue formation as well as severity of adhesion between scar tissue and the dura mater in a rat laminectomy model. Superficially, the peel-off test showed significantly decreased tenacity. In conclusion, the oxi-HA/ADH hydrogel is a promising injectable and thermosensitive material for prevention of post-operative epidural fibrosis.
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Adipatos/química , Materiales Biocompatibles/farmacología , Espacio Epidural/efectos de los fármacos , Ácido Hialurónico/química , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacología , Laminectomía/métodos , Animales , Materiales Biocompatibles/química , Línea Celular , Espacio Epidural/patología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibrosis/etiología , Fibrosis/prevención & control , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Laminectomía/efectos adversos , Ratones , Células 3T3 NIH , Oxidación-Reducción , Células PC12 , RatasRESUMEN
AIM: The aim was to evaluate the association of neoadjuvant therapy with increases in the incidence of anastomotic leakage (AL) after middle and low rectal anterior resection. METHOD: The electronic databases of PubMed, Web of Science, Scopus and Ovid were searched between 1980 and 2015. The random effects model was used to model the pooled data to determine the odds ratio with 95% confidence interval. Heterogeneity was evaluated using the Q test and I2 statistics. Subgroup, sensitivity and meta-regression analysis was conducted to explore heterogeneity. RESULTS: Neoadjuvant therapy was not shown to increase the incidence of postoperative AL as demonstrated by an OR of 1.16 [95% CI 0.99-1.36; P = 0.07 (random effects model)]. The subgroup analysis of neoadjuvant radiotherapy using the random effects model suggested that it did not increase the rate of postoperative AL (OR = 1.24, 95% CI 0.97-1.58; P = 0.08). The subgroup analysis of neoadjuvant chemoradiotherapy indicated that the rate of postoperative AL again did not increase with an OR = 1.06 [95% CI 0.86-1.30; P = 0.59 (random effects model)]. The interval to surgery after neoadjuvant therapy and preoperative radiotherapy (short or long course) was not associated with an increased incidence of postoperative AL. CONCLUSION: Neoadjuvant therapy does not appear to increase the incidence of postoperative AL after anterior resection for mid and low rectal cancer. In addition, neither the interval to surgery after neoadjuvant therapy nor the radiotherapy regimen increases the rate of postoperative AL.
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Anastomosis Quirúrgica/efectos adversos , Fuga Anastomótica/etiología , Quimioradioterapia/efectos adversos , Terapia Neoadyuvante/efectos adversos , Neoplasias del Recto/terapia , Adulto , Anciano , Fuga Anastomótica/epidemiología , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Factores de Riesgo , Factores de TiempoRESUMEN
Objective: To analyze the clinical features, diagnosis and differential diagnosis of peripheral primary neuroectodermal tumor(pPNET). Methods: The clinical data and diagnosis of a patient with rapid progressive pPNET in Shandong Provincial Hospital affiliated to Shandong University in January 2016 was reported and the related literatures were reviewed.The literature reviews were carried out respectively in CNKI, Wanfang and PubMed by July 2016 with "primitive neurotodermal tumour" and "PNET" being the search term from March 1994 to July 2016, including 13 articles. Results: A 41 year-old male patient was admitted to the hospital because of shortness of breath and occasional chest tightness, accompanied by general asthenia of about 15 d. Positron emission and transmission-CT of total trunk showed a mass in the right femoral osteoperiosteal mass, and multiple nodules in the left lung, lumps and nodules in the right lung, and right pleural thickening and effusion. Thoracoscopy was performed and pathology study confirmed the diagnosis of peripheral primary neuroectodermal tumor. The patient was given 2 courses of chemotherapy, but had rapid progressive worsening and died 1 month after PNET diagnosis. A total of 13 literatures of PNET were retrieved, all of which were case reports, and a total of 15 cases were reported.There were 9 male and 7 female patients, with a median age of 26 years.The symptoms had no specificity and most of them were solid masses in chest images, with or without pleural effusion. Eight cases were diagnosed by imaging guided percutaneous biopsy, 7 by operation and pathology, and 1 by medical thoracoscopy. Four cases underwent chemotherapy, and the survival time was less than 6 months. Twelve cases got surgical resection: 2 with surgical treatment, 7 with postoperative chemotherapy, 2 with postoperative radiotherapy and chemotherapy, 1 with postoperative chemotherapy and autologous bone marrow transplantation treatment, among which 11 patients completed follow-up. During follow-up, 3 cases died, and the survival time was 10 months, 3 years and 7 years, respectively. Conclusions: PNET is rare.Due to the lack of specificity of clinical manifestations, clinical diagnosis depends on biopsy. Early diagnosis and surgical resection are especially important for prognosis and quality of life.
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Biopsia , Tumores Neuroectodérmicos Periféricos Primitivos/patología , Adulto , Femenino , Humanos , Masculino , Pronóstico , Calidad de Vida , TóraxRESUMEN
OBJECTIVE: To investigate the clinical significance of the three-step approach in laparoscopic hemihepatectomy. METHODS: A total of 32 patients received laparoscopic hemihepatectomy with the three-step approach in Yijishan Hospital affiliated to Wannan Medical College between Aug 2013 and Oct 2015. All patients underwent thin slice CT scan and hemihepatectomy was imitated with the imagic explorer, preoperatively. The vessel distribution was observed at the section and the three-step approach was used in the hemihepatectomy. Pre- and post-operative data were collected and analyzed retrospectively. RESULTS: The length of middle hepatic vein (MHV) was (59.1±12.9) mm and the number of branchs to the left and right lobe were 3.07±0.78 and 3.11±0.64 respectively. The distance between the first branch of MHV and the diaphragmatic surface was (28.07±3.74) mm and the distance between MHV and the visceral surface was (14.4±4.3) mm. The laparoscopic surgeries (left hemihepatectomy in 28 and right hemihepatectomy in 4) were performed successfully in all cases with the three-step approach, without any conversion to the open surgeries. The operation time was (165±42) min in left hemihepatectomy and (305±50) min in right hemihepatectomy. The intraoperative blood loss was (242±65) ml in left hemihepatectomy and (695±122)ml in right hemihepatectomy. All the patients recovered well without severe complications except for bile leakage in 3 patients who were cured with drainage. The postoperative hospital stay was (7.96±1.8) d. CONCLUSIONS: the distribution of vessel is regional at the section of hemihepatectomy and the three-step approach based on this feature is safe and effective in laparoscopic hemihepatectomy, which can shorten the operation time and reduce the difficulty of operation.
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Hepatectomía/métodos , Venas Hepáticas/diagnóstico por imagen , Laparoscopía/métodos , Hígado/cirugía , Pérdida de Sangre Quirúrgica , Hepatectomía/instrumentación , Humanos , Laparoscopía/instrumentación , Tiempo de Internación , Hígado/diagnóstico por imagen , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
An increase in interleukin (IL)-17A-producing cells, particularly at sites of tissue inflammation, is observed frequently, yet the mechanism is not fully understood. This study aims to dissect the role of IL-17 in autoimmunity-mediated neuroinflammation. The cytokine milieu containing elevated IL-17, which often appears in active states of autoimmunity, was mimicked in vitro by a supernatant obtained from rat peripheral blood monocytes stimulated with phorbol mystistate acetate (PMA)/ionomycin. The application of such inflammatory media on only primary cultured cerebellar granule neurones resulted in significant apoptosis, but the presence of astrocytes largely prevented the effect. The supernatants of the stimulated astrocytes, especially those that contained the highest level of IL-17, achieved the best protection, and this effect could be blocked by anti-IL-17 antibodies. Protein IL-17 inhibited intracellular calcium increase and protected the neurones under inflammatory attack from apoptosis. IL-17, but not interferon (IFN)-γ, in the inflammatory media contributed to astrocyte secretion of IL-17, which depended on the Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway activation. The astrocytes that were treated with IL-17 alone or with prolonged treatment of the inflammatory media failed to produce sufficient levels of IL-17. Moreover, confirmatory data were obtained in vivo in a monophasic experimental autoimmune uveitis (EAU) in Lewis rats; in this preparation, the high-level IL-17-containing the cytokine milieu was demonstrated, along with IL-17 secretion by the resident neural cells. The antagonism of IL-17 at a late stage disturbed the disease resolution and resulted in significant neural apoptosis. Our data show a dynamic role of IL-17 in the maintenance of homeostasis and neuroprotection in active neuroinflammation.
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Astrocitos/inmunología , Inflamación/inmunología , Interleucina-17/metabolismo , Neuronas/inmunología , Animales , Anticuerpos Bloqueadores/inmunología , Apoptosis/inmunología , Astrocitos/metabolismo , Autoinmunidad/inmunología , Interferón gamma/metabolismo , Interleucina-17/inmunología , Quinasas Janus/metabolismo , Leucocitos Mononucleares/efectos de los fármacos , Neuronas/metabolismo , ARN Mensajero/biosíntesis , Ratas , Ratas Endogámicas Lew , Ratas Wistar , Factores de Transcripción STAT/metabolismo , Acetato de Tetradecanoilforbol , Uveítis/inmunologíaRESUMEN
OBJECTIVE: The aim of this study was to observe clinical curative effects of combination application of dimethylbiguanide and pioglitazone and single application of pioglitazone in patients with polycystic ovarian syndrome (PCOS) complicated with insulin resistance (IR). MATERIALS AND METHODS: Forty cases of patients with PCOS complicated with IR were investigated, and 20 cases of infertile women without PCOS were taken as the control group. PCOS group was divided into group A and group B according to body mass index (BMI) to detect glucose and lipids metabolism indicators, C reactive protein (CRP), etc. There were 20 cases in group A (Pioglitazone) and 20 cases in group B (dimethylbiguanide and pioglitazone). After treatment for 12 weeks, changes of the above various indicators were compared. RESULTS: After treatment, insulin resistance index and serum testosterone (T) of two groups patients with PCOS significantly reduced (p < 0.05). Compared to before treatment, BMI of group B significantly reduced (p < 0.05). For INS at two hours after treatment, group B reduced more significantly (p < 0.05). CONCLUSION: The combination of dimethylbiguanide and pioglitazone was more effective for the treatment of PCOS complicated with IR than simple pioglitazone; chronic inflammation occurrence was possibly one of reasons for insulin sensitivity reduction of patients with PCOS.
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Hiperinsulinismo/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Resistencia a la Insulina , Metformina/uso terapéutico , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Tiazolidinedionas/uso terapéutico , Adolescente , Adulto , Glucemia/metabolismo , Índice de Masa Corporal , Proteína C-Reactiva/metabolismo , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Quimioterapia Combinada , Ayuno , Femenino , Homeostasis , Humanos , Hiperinsulinismo/sangre , Hiperinsulinismo/complicaciones , Infertilidad Femenina/sangre , Insulina/sangre , Obesidad/sangre , Obesidad/complicaciones , Pioglitazona , Síndrome del Ovario Poliquístico/sangre , Síndrome del Ovario Poliquístico/complicaciones , Estudios Prospectivos , Testosterona/sangre , Triglicéridos/sangre , Adulto JovenRESUMEN
Objective: Analysis of the characteristics of influenza epidemic in Anhui Province and quantification of the impact of different factors on influenza occurrence, providing scientific basis for better influenza prevention and control. Methods: Descriptive analysis and factor analysis were conducted on influenza-like illness (ILI) cases and RT-PCR results in Anhui Province from 2013 to 2021 using data from China's Influenza Monitoring Information System. Results: The percentage of influenza-like illness (ILI%) of sentinel hospitals in Anhui Province from April 1, 2013 to March 31, 2021 was 3.80% (1 209 142/31 779 987), showing an overall increasing trend, with a relatively high proportion in 2017-2018 at 4.30% (191 148/4 448 211). The proportion of ILI cases in infants and young children aged 0-4 years was a relatively high at 54.14% (654 676/1 209 142), and the highest ILI% was observed in Fuyang City, Anhui Province (6.25%, 236 863/3 788 863). Laboratory monitoring results showed that the positive rate of ILI cases in sentinel hospitals in 8 influenza monitoring years was 16.38% (34 868/212 912), showing an increasing trend year by year, with a relatively proportion in 2017-2018 at 26.19% (6 936/26 488). The detection rate of school-age children aged 5-14 years was a relativelyhigh at 28.81% (13 869/48 144), and the positive rate was a relatively high in Wuhu City among the 16 cities, reaching 22.01% (2 693/122 237). Influenza activity showed a single peak in winter-spring and alternating double peaks in winter-spring and summer, with different subtypes alternating, and A (H3N2) was the dominant subtype in summer. The results of a multiple logistic regression model showed that the positive rate was higher in 2017-2018, among children aged 5-14 years, in winter, and in southern Anhui. Conclusions: Influenza epidemic in Anhui Province has a clear seasonal pattern, and the ILI% and detection rate have shown an upward trend from 2013 to 2021. Therefore, it is suggested to ensure vaccine supply before the winter-spring influenza season arrives, and to strengthen vaccine uptake and health education to avoid the risk of infection during the peak period of influenza.
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Vacunas contra la Influenza , Gripe Humana , Niño , Lactante , Humanos , Preescolar , Gripe Humana/epidemiología , Subtipo H3N2 del Virus de la Influenza A , Ciudades , Factores de RiesgoRESUMEN
Objective: Continuous glucose monitoring (CGM) were performed in children with hepatic glycogen storage disease (GSD) to accurately understand the situation of glucose levels during their treatment, and to provide support for optimizing their nutritional management. Methods: In this retrospective research, 42 patients with hepatic GSD who under went 72 h CGM were collected from Department of Pediatrics, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology from October 2019 to January 2020. According to the genetic test results, they were divided into 5 groups: type â a, type â b, type â ¢a, type â ¥ and type â ¨a. After long-term follow up and regular treatment, the clinical data (induding course, age, height, weight and biochemical parameters, etc.) on the day of CGM were summarized, and 72 h CGM were performed to assess the occurrence of hypoglycemia and hyperglycemia.χ² test, Fisher exact probability method, t test, analysis of variance or nonparametric test were used for comparison between groups. Results: Forty-two cases of hepatic GSD patients included 25 males and 17 females (20 cases of type â a, 3 cases of type â b, 10 cases of type â ¢a, 3 cases of type â ¥ and 6 cases of type â ¨a).The age was 9.5 (6.7, 12.9) years, and the course of disease was 6.8 (5.1, 11.3) years. The average levels of glucose of the patients were all normal. However, the levels of standard deviation of blood glucose (SDBG) and mean amplitude of glycemic excursion (MAGE) were significantly different (F=2.747, 3.029,both P<0.05). Among them, the SDBG of type â a and â ¢a were significantly higher than those of type â ¨a ((1.10±0.36), (0.98±0.30) vs. (0.62±0.26) mmol/L, t=3.010, 2.440, both P<0.05), while the MAGE of type â was higher than that of â ¨a and â ¢a ((2.3±0.9) mmol/L vs. (1.2±0.6) and (1.7±0.6) mmol/L, t=2.734, 2.302, both P<0.05, respectively). Conclusions: CGMS can accurately assess the fluctuations of blood glucose and effectively detect hidden hypoglycemia and hyperglycemia in hepatic GSD patients. For different types of hepatic GSD, individualized corn starch treatment doses should be given according to the different situation of blood glucose, so as to optimize the patient's treatment and improve their prognosis.
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Glucemia , Enfermedad del Almacenamiento de Glucógeno , Automonitorización de la Glucosa Sanguínea , Niño , Femenino , Glucosa , Humanos , Masculino , Estudios RetrospectivosRESUMEN
The effects of delayed first feeding on growth and survival of spotted mandarin fish Siniperca scherzeri larvae were examined under controlled conditions. Morphometric characters [yolk-sac volume, oil globule volume, head depth (H(D)), body depth (B(D)), eye diameter (E(D)), musculature height (M(H)), mouth diameter (M(D)) and total length (L(T))], body mass (M), specific growth rate (S(GR)) and survival were evaluated under different first-feeding time (2, 3, 4 and 5 days after hatching). Larvae began to feed exogenously at 2 days after hatching (DAH) and the point of no return (P(NR)) occurred between 5 and 6 DAH at 23 degrees C, range +/-1.0 degrees C. The yolk volume of larvae first-fed at 2 days had a significant difference compared with that of larvae first-fed at 3, 4 and 5 days on 3 and 4 DAH. The larvae first-fed at 2 days achieved comparatively better growth performance than that of 3, 4 and 5 days. On 5 DAH, all morphometric characters had significant differences between 2 and 5 days and 2 and 4 days initial feeding, respectively. Total mortality was recorded on 9 DAH for the larvae first-fed at 5 days. On 12 DAH, significant differences were observed between 2 and 4 days and 3 and 4 days initial feeding for all morphometric characters. From 16 DAH to the end of experiment, all growth variables of the larvae first-fed at 2 days were significantly higher than those in other treatments. The S(GR) (2-9 DAH) first-fed at 2 and 3 days were significantly higher than 4 and 5 day treatments, and the S(GR) (9-16 DAH) first-fed at 2 days was significantly higher than 3 and 4 day treatments. There was no significant difference, however, of S(GR) (16-28 DAH) among treatments. Survival rate was significantly higher at 2 days initial feeding (27.42%) when compared with 3 (15.96%) and 4 days (7.92%) initial feeding at the end of experiment. The present study suggests that the first feeding of S. scherzeri larvae should be initiated at 2 days after hatching for achieving good growth and survival.
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Métodos de Alimentación/veterinaria , Explotaciones Pesqueras/métodos , Perciformes/fisiología , Animales , Perciformes/crecimiento & desarrollo , Análisis de Supervivencia , Factores de Tiempo , Saco Vitelino/metabolismoRESUMEN
Comfort eating during periods of stress is a common phenomenon observed in both animals and humans. However, the underlying mechanisms of stress-induced food intake remain elusive. The amygdala plays a central role in higher-order emotional processing and the posterodorsal subnucleus of the medial amygdala (MePD), in particular, is involved in food intake. Extra-hypothalamic corticotrophin-releasing factor (CRF) is well recognised for mediating behavioural responses to stress. To explore the possible role of amygdala CRF receptor activation in stress-induced food intake, we evaluated whether a stressor such as tail-pinch, which reliably induces food intake, would fail to do so in animals bearing bilateral neurotoxic lesions of the MePD. Our results showed that ibotenic acid induced lesions of the MePD markedly reduced tail-pinch induced food intake in ovariectomised, 17ß-oestradiol replaced rats. In addition, intra-MePD (right side only) administration of CRF (0.002 or 0.02 ng) via chronically implanted cannulae resulted in a dose-dependent increase in food intake, although higher doses of 0.2 and 2 ng CRF had less effect, producing a bell shaped curve. Furthermore, intra-MePD (bilateral) administration of the CRF receptor antagonist, astressin (0.3 µg per side) effectively blocked tail-pinch induced food intake. These data suggest that the MePD is involved in stress-induced food intake and that the amygdala CRF system may be a mediator of comfort eating.
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Complejo Nuclear Corticomedial/fisiopatología , Ingestión de Alimentos , Estrés Psicológico/fisiopatología , Animales , Complejo Nuclear Corticomedial/efectos de los fármacos , Hormona Liberadora de Corticotropina/administración & dosificación , Ingestión de Alimentos/efectos de los fármacos , Estradiol/administración & dosificación , Femenino , Ovariectomía , Fragmentos de Péptidos/administración & dosificación , Estimulación Física , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidoresRESUMEN
The role of glycosylation on the enzymatic properties of single chain urokinase-type plasminogen activator (scu-PA) was investigated by site-specific mutagenesis of the glycosylated Asn-302 residu to Gln. In addition, the role of the NH2-terminal polypeptide chain and of the Cys-148 to Cys-279 interchain disulphide bond on the activity of non-glycosylated scu-PA was investigated. Therefore, variants of recombinant scu-PA (rscu-PA) were produced by transfecting Chinese hamster ovary cells with cDNA encoding rscu-PA N302Q (rscu-PA with Asn-302 to Gln mutation), rscu-PA C279A,N302Q (rscu-PA with Cys-279 to Ala and Asn-302 to Gln mutations) or rscu-PA del(N2-F157)C279A,N302Q (rscu-PA C279A,N302Q with deletion of Asn-2 through Phe-157). These mutants were purified to homogeneity from conditioned cell culture medium and were obtained essentially as single chain molecules with specific activities on fibrin plates of (mean +/- S.E.; n = 6) 45,000 +/- 5000. IU/mg, 19,000 +/- 800 IU/mg and < or = 100 IU/mg for rscu-PA N302Q, rscu-PA C279A,N302Q and rscu-PA del(N2-F157)C279A,N302Q, respectively, as compared to 64,000 +/- 2600 IU/mg for wild-type rscu-PA obtained in the same expression system. Plasmin quantitatively converts rscu-PA N302Q and rscu-PA C279A,N302Q to amidolytically active two-chain derivatives with a specific activity of 56,000 IU/mg and 32,000 IU/mg, respectively, as compared to 75,000 IU/mg for wild-type rscu-PA. Plasminogen activation as a function of time was comparable for rscu-PA N302Q and wild-type rscu-PA, and somewhat slower for rscu-PA C279A,N302Q. In a human plasma milieu in vitro, consisting of a 125I-fibrin labeled plasma clot submerged in plasma, 50 percent clot lysis in 2 h required 2.2 micrograms/ml rscu-PA N302Q and 6.0 micrograms/ml rscu-PA C279A,N302Q, as compared to 3.2 micrograms/ml wild-type rscu-PA. In contrast, rscu-PA del(N2-F157)C279A,N302Q was not converted to an amidolytically active two chain derivative by plasmin, and did not induce significant plasminogen activation in purified systems or clot lysis in a human plasma milieu. Following bolus injections in hamsters, the initial half-lives (1.8-2.6 min) and the plasma clearances (0.6-1.5 ml min-1) were comparable for wild-type rscu-PA and for the three rscu-PA mutants. These results suggest that the fibrinolytic activity in a plasma milieu in vitro and the in vivo turnover of rscu-PA are not markedly affected by the absence of carbohydrate.(ABSTRACT TRUNCATED AT 400 WORDS)
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Activador de Plasminógeno de Tipo Uroquinasa/química , Animales , Secuencia de Bases , Cricetinae , Activación Enzimática , Fibrinolisina/farmacología , Fibrinólisis , Glicosilación , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Oligodesoxirribonucleótidos/química , Plasminógeno/metabolismo , Proteínas Recombinantes , Relación Estructura-Actividad , Activador de Plasminógeno de Tipo Uroquinasa/metabolismo , Activador de Plasminógeno de Tipo Uroquinasa/farmacocinéticaRESUMEN
Obesity is the major risk factor for early puberty, but emerging evidence indicates other factors including psychosocial stress. One key brain region notable for its role in controlling calorie intake, stress, and behavior is the amygdala. Early studies involving amygdala lesions that included the medial nucleus advanced puberty in rats. More recently it was shown that a critical site for lesion-induced hyperphagia and obesity is the posterodorsal subnucleus of the medial amygdala (MePD), which may explain the advancement of puberty. Glutamatergic activity also increases in the MePD during puberty without a corresponding γ-aminobutyric acid (GABA)ergic change, suggesting an overall activation of this brain region. In the present study, we report that neurotoxic lesioning of the MePD advances puberty and increases weight gain in female rats fed a normal diet. However, MePD lesioned rats fed a 25% nonnutritive bulk diet also showed the dramatic advancement of puberty but without the increase in body weight. In both dietary groups, MePD lesions resulted in an increase in socialization and a decrease in play fighting behavior. Chronic GABAA receptor antagonism in the MePD from postnatal day 21 for 14 days also advanced puberty, increased socialization, and decreased play fighting without altering body weight, whereas glutamate receptor antagonism delayed puberty and decreased socialization without affecting play fighting. In conclusion, our results suggest the MePD regulates the timing of puberty via a novel mechanism independent of change in body weight and caloric intake. MePD glutamatergic systems advance the timing of puberty whereas local GABAergic activation results in a delay.
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Amígdala del Cerebelo/fisiología , Maduración Sexual/fisiología , Conducta Social , Aumento de Peso/fisiología , Amígdala del Cerebelo/metabolismo , Animales , Bicuculina/farmacología , Ciclo Estral/fisiología , Femenino , Antagonistas de Receptores de GABA-A/farmacología , Embarazo , Ratas Sprague-Dawley , Receptores de GABA-A/metabolismo , Maduración Sexual/efectos de los fármacos , Factores de Tiempo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Kisspeptin plays a critical role in pubertal timing and reproductive function. In rodents, kisspeptin perikarya within the hypothalamic arcuate (ARC) and anteroventral periventricular (AVPV) nuclei are thought to be involved in LH pulse and surge generation, respectively. Using bilateral microinjections of recombinant adeno-associated virus encoding kisspeptin antisense into the ARC or AVPV of female rats at postnatal day 10, we investigated the relative importance of these two kisspeptin populations in the control of pubertal timing, estrous cyclicity, and LH surge and pulse generation. A 37% knockdown of kisspeptin in the AVPV resulted in a significant delay in vaginal opening and first vaginal estrous, abnormal estrous cyclicity, and reduction in the occurrence of spontaneous LH surges, although these retained normal amplitude. This AVPV knockdown had no effect on LH pulse frequency, measured after ovariectomy. A 32% reduction of kisspeptin in the ARC had no effect on the onset of puberty but resulted in abnormal estrous cyclicity and decreased LH pulse frequency. Additionally, the knockdown of kisspeptin in the ARC decreased the amplitude but not the incidence of LH surges. These results might suggest that the role of AVPV kisspeptin in the control of pubertal timing is particularly sensitive to perturbation. In accordance with our previous studies, ARC kisspeptin signaling was critical for normal pulsatile LH secretion in female rats. Despite the widely reported role of AVPV kisspeptin neurons in LH surge generation, this study suggests that both AVPV and ARC populations are essential for normal LH surges and estrous cyclicity.
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Núcleo Arqueado del Hipotálamo/metabolismo , Ciclo Estral/genética , Hipotálamo Anterior/metabolismo , Kisspeptinas/genética , Neuronas/metabolismo , Pubertad/genética , Maduración Sexual/genética , Animales , Núcleo Arqueado del Hipotálamo/citología , Ciclo Estral/metabolismo , Femenino , Técnicas de Silenciamiento del Gen , Hipotálamo Anterior/citología , Kisspeptinas/metabolismo , Hormona Luteinizante/metabolismo , Neuronas/citología , Pubertad/metabolismo , RatasRESUMEN
AIM: To explore the pathomorphological characteristics of large hepatocellular carcinoma (LHCC) with low serum alpha-fetoprotein (AFP) level. METHODS: Specimens obtained from surgically resected LHCC with undetectable or low levels of serum AFP were fixed in formalin, embedded in paraffin, and prepared as serial sections. Routine hematoxylin and eosin as well as immunohistochemical stains (LSAB method) were used to test for expression of AFP, alpha-1-antitrypsin, epithelial membrane antigen, and vimentin. Some characteristics of the histopathological changes and immunohistochemical reactions of the cancerous tissues were observed under the light microscope. RESULTS: The majority of the cases (19/30) of LHCC with undetectable or low levels of serum AFP were of the clear-cell-type HCC, with 2 being positive for AFP expression at the periphery of the cytoplasm. CONCLUSION: The clear cell is the morphological manifestation of disturbance in glycogen and/or lipid metabolism of hepatoma cells. Such changes might be one of the factors hindering the synthesis of AFP and resulting in negative or low level serum AFP of the patient.
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The construction of a gene encoding Lys-human proinsulin, its direct expression in E. coli, and the simple purification procedure are described here. The temperature inducible promotor was employed for induction in a very short time. The expression level could reach 20-30%. After simple downstream processing and only one step of Sephadex G50 purification, 150 mg recombinant Lys-human proinsulin with a purity of up to 90% could be obtained easily from 1 L of high density fermentation medium. The obtained product is in the form of Met-Lys-human proinsulin because of the failure of the bacterial host to remove the initiator methionine residue. The Lys-human proinsulin could be changed into human insulin by trypsin and carboxypeptidase B treatment in later steps. After separation with DEAE-Sephadex A25, human insulin with expected amino acid composition and full native biological activity could be obtained with a yield of 50 mg/L of fermentation medium.
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Insulina/biosíntesis , Proinsulina/genética , Secuencia de Bases , Escherichia coli/genética , Vectores Genéticos , Humanos , Insulina/genética , Datos de Secuencia Molecular , Plásmidos , Proinsulina/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Análisis de SecuenciaRESUMEN
The P68 nuclear protein is an ATP-dependent RNA helicase and RNA-dependent ATPase, and has been proved to have functions in the regulation of cell growth and division. Different phenotypes of P68 between tumouric cell lines (HeLa, TC3H10) and non-tumouric cell lines (NIH3T3 and NC3H10) were observed, and we reasoned that overexpression of P68 might profoundly affect cellular growth properties. We report here that overexpression human P68 by transfection of recombinant P68 expression vector in NIH3T3 and NC3H10 fibroblasts causes their tumorigenic transformation as determined by four criteria: (1) the morphology of transfectants from long fibric to spindle shaped; (2) formation of transformed foci on the monolayer of cells; (3) the transfectants acquired the ability to grow on soft agar forming cell microsphere cube; (4) tumor formation in nude mice after injection of transfected cells for 3-4 weeks.
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Transformación Celular Neoplásica , Proteínas Quinasas/fisiología , ARN Helicasas/fisiología , Células 3T3 , Animales , ARN Helicasas DEAD-box , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Neoplasias Experimentales/etiología , TransfecciónRESUMEN
The neural mechanisms controlling puberty onset remain enigmatic. Humans with loss of function mutations in TAC3 or TACR3, the genes encoding neurokinin B (NKB) or its receptor, neurokinin-3 receptor (NK3R), respectively, present with severe congenital gonadotrophin deficiency and pubertal failure. Animal studies have shown ambiguous actions of NKB-NK3R signalling with respect to controlling puberty onset. The present study aimed to determine the role of endogenous NKB-NK3R signalling in the control of pulsatile luteinising hormone (LH) secretion and the timing of puberty onset, and also whether precocious pubertal onset as a result of an obesogenic diet is similarly regulated by this neuropeptide system. Prepubertal female rats, chronically implanted with i.c.v. cannulae, were administered SB222200, a NK3R antagonist, or artificial cerebrospinal fluid via an osmotic mini-pump for 14 days. SB222200 significantly delayed the onset of vaginal opening and first oestrus (as markers of puberty) compared to controls in both normal and high-fat diet fed animals. Additionally, serial blood sampling, via chronic indwelling cardiac catheters, revealed that the increase in LH pulse frequency was delayed and that the LH pulse amplitude was reduced in response to NK3R antagonism, regardless of dietary status. These data suggest that endogenous NKB-NK3R signalling plays a role in controlling the timing of puberty and the associated acceleration of gonadotrophin-releasing hormone pulse generator frequency in the female rat.
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Hormona Luteinizante/sangre , Neuroquinina B/fisiología , Pubertad/efectos de los fármacos , Pubertad/fisiología , Receptores de Neuroquinina-3/antagonistas & inhibidores , Receptores de Neuroquinina-3/fisiología , Animales , Dieta Alta en Grasa , Femenino , Hormona Luteinizante/efectos de los fármacos , Masculino , Microinyecciones , Quinolinas/administración & dosificación , Quinolinas/farmacología , RatasRESUMEN
Acute systemic stress disrupts reproductive function by inhibiting pulsatile gonadotropin secretion. The underlying mechanism involves stress-induced suppression of the GnRH pulse generator, the functional unit of which is considered to be the hypothalamic arcuate nucleus kisspeptin/neurokinin B/dynorphin A neurons. Agonists of the neurokinin B (NKB) receptor (NK3R) have been shown to suppress the GnRH pulse generator, in a dynorphin A (Dyn)-dependent fashion, under hypoestrogenic conditions, and Dyn has been well documented to mediate several stress-related central regulatory functions. We hypothesized that the NKB/Dyn signaling cascade is required for stress-induced suppression of the GnRH pulse generator. To investigate this ovariectomized rats, iv administered with Escherichia coli lipopolysaccharide (LPS) following intracerebroventricular pretreatment with NK3R or κ-opioid receptor (Dyn receptor) antagonists, were subjected to frequent blood sampling for hormone analysis. Antagonism of NK3R, but not κ-opioid receptor, blocked the suppressive effect of LPS challenge on LH pulse frequency. Neither antagonist affected LPS-induced corticosterone secretion. Hypothalamic arcuate nucleus NKB neurons project to the paraventricular nucleus, the major hypothalamic source of the stress-related neuropeptides CRH and arginine vasopressin (AVP), which have been implicated in the stress-induced suppression of the hypothalamic-pituitary-gonadal axis. A separate group of ovariectomized rats was, therefore, used to address the potential involvement of central CRH and/or AVP signaling in the suppression of LH pulsatility induced by intracerebroventricular administration of a selective NK3R agonist, senktide. Neither AVP nor CRH receptor antagonists affected the senktide-induced suppression of the LH pulse; however, antagonism of type 2 CRH receptors attenuated the accompanying elevation of corticosterone levels. These data indicate that the suppression of the GnRH pulse generator by acute systemic stress requires hypothalamic NKB/NK3R signaling and that any involvement of CRH therewith is functionally upstream of NKB.
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Hormona Liberadora de Gonadotropina/metabolismo , Neuroquinina B/metabolismo , Receptores de Neuroquinina-3/metabolismo , Transducción de Señal/fisiología , Animales , Antagonistas de los Receptores de Hormonas Antidiuréticas , Núcleo Arqueado del Hipotálamo/citología , Núcleo Arqueado del Hipotálamo/efectos de los fármacos , Núcleo Arqueado del Hipotálamo/metabolismo , Corticosterona/sangre , Corticosterona/metabolismo , Hormona Liberadora de Corticotropina/farmacología , Femenino , Inyecciones Intraventriculares , Lipopolisacáridos/administración & dosificación , Lipopolisacáridos/farmacología , Hormona Luteinizante/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Ovariectomía , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/farmacología , Pirimidinas/farmacología , Pirroles/farmacología , Quinolinas/administración & dosificación , Quinolinas/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Receptores de Neuroquinina-3/agonistas , Receptores de Neuroquinina-3/antagonistas & inhibidores , Receptores de Vasopresinas/metabolismo , Reproducción/fisiología , Transducción de Señal/efectos de los fármacos , Estrés Fisiológico/fisiología , Sustancia P/administración & dosificación , Sustancia P/análogos & derivados , Sustancia P/farmacologíaRESUMEN
Prolonged exposure to environmental stress activates the hypothalamic-pituitary-adrenal (HPA) axis and generally disrupts the hypothalamic-pituitary-gonadal axis. Because CRF expression in the central nucleus of the amygdala (CeA) is a key modulator in adaptation to chronic stress, and central administration of CRF inhibits the hypothalamic GnRH pulse generator, we tested the hypothesis that overexpression of CRF in the CeA of female rats alters anxiety behavior, dysregulates the HPA axis response to stress, changes pubertal timing, and disrupts reproduction. We used a lentiviral vector to increase CRF expression site specifically in the CeA of preweaning (postnatal day 12) female rats. Overexpression of CRF in the CeA increased anxiety-like behavior in peripubertal rats shown by a reduction in time spent in the open arms of the elevated plus maze and a decrease in social interaction. Paradoxically, puberty onset was advanced but followed by irregular estrous cyclicity and an absence of spontaneous preovulatory LH surges associated with proestrous vaginal cytology in rats overexpressing CRF. Despite the absence of change in basal corticosterone secretion or induced by stress (lipopolysaccharide or restraint), overexpression of CRF in the CeA significantly decreased lipopolysaccharide, but not restraint, stress-induced suppression of pulsatile LH secretion in postpubertal ovariectomized rats, indicating a differential stress responsivity of the GnRH pulse generator to immunological stress and a potential adaptation of the HPA axis to chronic activation of amygdaloid CRF. These data suggest that the expression profile of this key limbic brain CRF system might contribute to the complex neural mechanisms underlying the increasing incidence of early onset of puberty on the one hand and infertility on the other attributed to chronic stress in modern human society.